The structure of cyanine 5 terminally attached to double-stranded DNA: Implications for FRET studies

被引：84

作者：

Iqbal, Asif ^{[1
]}

Wang, Lihua ^{[1
]}

Thompson, Katherine C. ^{[2
]}

Lilley, David M. J. ^{[1
]}

Norman, David G. ^{[1
]}

机构：

[1] Univ Dundee, Canc Res UK Nucle Acid Struct Res Grp, Dundee DD1 5EH, Scotland

[2] Univ London Birkbeck Coll, Sch Biol & Chem Sci, London WC1E 7HX, England

来源：

BIOCHEMISTRY | 2008年 / 47卷 / 30期

基金：

英国生物技术与生命科学研究理事会;

关键词：

D O I：

10.1021/bi800773f

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Fluorescence resonance energy transfer, FRET, can be used to obtain long-range distance information for macromolecules and is particularly powerful when used in single-molecule studies. The determination of accurate distances requires knowledge of the fluorophore position with respect to the macromolecule. In this study we have used NMR to determine the structure of the commonly used fluorophore indocarbocyanine-5 (Cy5) covalently attached to the 5'-terminus of double-helical DNA. We find that Cy5 is predominantly stacked onto the end of the duplex, in a manner similar to an additional base pair. This is very similar to the behavior of Cy3 terminally attached to DNA and suggests that the efficiency of energy transfer between Cy3 and Cy5, that are attached to nucleic acids in this way, will exhibit significant dependence on fluorophore orientation.

引用

页码：7857 / 7862

页数：6

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